JP5912181B2 - Cell tissue inspection system, cell culture device, and cell tissue inspection method - Google Patents

Description

  The present invention relates to a cell tissue inspection system, a cell culture apparatus, and a cell tissue inspection method, and more particularly to a system and method for non-invasively inspecting cell quality.

  In regenerative medicine, which uses his own cells or other people's cells to treat diseases, cells collected from living organisms are often treated after culturing to increase the number of cells or to form tissues in an appropriate shape. Used. Culture of cells used for treatment must be performed in accordance with GMP (Good Manufacturing Practice) in a cell culture center called Cell Processing Center (CPC). For example, in the case of a tissue that is directly transplanted to an affected part such as a cell sheet, the quality evaluation is often performed by analyzing an evaluation sample prepared simultaneously with the graft, and the transplanted tissue itself is not evaluated. Currently. In addition, the degree of cell proliferation during the culture is evaluated only by the current phase contrast microscope, and it takes time when multiple points need to be observed.

  As means for solving such problems, for example, Non-Patent Documents 1 and 2 disclose means for measuring the barrier function of epithelial cells and measuring the degree of cell proliferation non-invasively by measuring electrical resistance. By applying a weak alternating current or alternating voltage, the frequency-dependent impedance composed of resistance (R: resistance) and capacitance (C: capacitance) is measured. It is possible to calculate an increase in resistance and an increase in impedance and a decrease in capacitance due to an increase in the number of adherent / extended cells, and devices equipped with such means have been developed. Further, for example, Patent Documents 1 and 2 disclose measuring means for measuring cell activity (for example, respiratory activity), tissue characteristics (for example, action potential), and the like. Non-invasive measuring means can be applied to quality inspection.

JP 2010-261923 A JP 2011-89894 A

J. Wegener et al., “Automated multi-well device to measure transepithelial electrical resistances under physiological conditions”, BioTechniques, Vol.37, No.4 (2004), pp.590-596 Solly K., “Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays”, Assay Drug Dev. Technol., Vol. 2 (4), 363-372 (2004)

  Regarding the examination of cell quality, there is a need to evaluate various indicators such as the degree of cell proliferation and cell activity during culture using various non-invasive measurement means depending on the cell / tissue type.

  However, the techniques disclosed in Patent Documents 1 and 2 and Non-Patent Documents 1 and 2 are specialized for examinations of specific applications and purposes, and in cell culture, quality evaluation is automatically performed using multiple indices. There is no mention of that. That is, conventionally, in a cell culture apparatus that automatically cultivates cells and creates tissue, each non-invasive measurement means is organically linked, and the measurement by the next measurement means is performed based on the measurement result by a certain measurement means. There is no cell culture device and system that can automatically instruct quality evaluation based on a plurality of indices.

  The object of the present invention is to solve these problems and perform a series of multi-step quality inspections necessary for quality evaluation of cells / tissues with a plurality of indices, in a non-invasive and automatic manner, a cell culture device, a cell tissue inspection It is to provide a system and a cell tissue examination method.

  An example of a representative example of the present invention is as follows. A cell tissue inspection system for inspecting a tissue of a cell, comprising a measurement device used for the inspection, a quality inspection unit that controls inspection of a cell tissue to be measured using the measurement device, and information relating to the inspection A database for holding, the test includes at least a test for grasping a growth state of the cell, measures an impedance of the cell tissue to be measured, and compares and determines the measurement result with a predetermined first threshold value. And the contents of the next examination are changed depending on whether the threshold value is equal to or greater than the first threshold value or not.

  According to the present invention, it is possible to perform quality inspection using a plurality of indices of cells / tissues to be transplanted non-invasively and automatically, thereby realizing safe and secure regenerative medicine.

It is a figure which shows the structural example of the cell culture apparatus based on 1st Example of this invention. It is a figure which shows the example of the quality inspection program based on 1st Example. It is a figure explaining the relation between cell density and high frequency impedance concerning the 1st example. It is a figure explaining the relationship between a cell seeding density | concentration and an impedance based on a 1st Example. It is a figure explaining the relationship between cell activity and a respiratory activity value based on a 1st Example. It is a figure explaining the relation between barrier functionality and the low frequency impedance concerning the 1st example. It is a figure explaining the layering degree understood by the cell and tissue shape measurement based on a 1st Example. It is a figure which shows the example of the display screen in a 1st Example. It is a figure which shows the example of the quality inspection program based on 2nd Example of this invention. It is a figure explaining the relation between action potential and membrane potential value concerning the 2nd example. It is a figure which shows the example of the quality inspection program based on 3rd Example of this invention. It is a figure which shows the other example of the display screen based on the 4th Example of this invention.

In the present invention, there is provided a cell tissue examination system that monitors impedance over time during culture and changes the next examination content according to the presence or absence of the change.
INDUSTRIAL APPLICABILITY The present invention is useful as a cell culture device that realizes quality inspection using a plurality of noninvasive cell indices.

  Various embodiments of the present invention will be described below with reference to the accompanying drawings. However, these examples are merely examples for realizing the present invention, and do not limit the technical scope of the present invention. In each drawing, the same reference numerals are assigned to common components.

A cell tissue examination system according to a first embodiment of the present invention will be described with reference to FIGS. FIG. 1 is a block diagram for explaining a functional configuration of the cell culture device 1 according to the first embodiment. The cell culture device 1 includes a control device 2, a thermostatic chamber 3 in which a cell culture container 4 is disposed, and each component controlled by the control device 2. The cell culture container 4 has, for example, a two-layer structure in which an upper layer culture container for culturing a regenerated tissue is accommodated in a lower layer culture container for culturing cells. Each component of the cell culture device 1 controlled by the control device 2 is appropriately connected to the thermostat 3 and the cell culture container 4 disposed inside the thermostat. The control device 2 includes a temperature adjusting unit 5 for controlling the temperature of the thermostat 3, a humidity adjusting unit 6 for controlling the humidity in the culture vessel, and a gas concentration in the culture vessel, A gas concentration adjusting unit 8 having a gas supply unit 7 and a culture solution supply pump having a liquid feeding tube connected to a tank 9 for holding the culture solution and waste solution for automatically exchanging the culture solution in the culture vessel 10, temperature / humidity / CO ₂ / O ₂ sensor 11, cell observation device 12 such as a camera for the purpose of confirming the cell morphology in the cell culture container 4, cell proliferation status and barrier measurement The impedance measurement device 13, the cell / tissue shape measurement device 14, the action potential measurement device 15, the cell activity measurement device 16, and a display 18 that displays a display screen 17 as a user interface are connected. To have. Each of the measuring devices is incorporated in advance in the cell culture container 4 to be measured, and can measure the state of the cells being cultured in the culture container non-invasively. For example, the impedance measuring device 13 can monitor the impedance of cells in culture over time. Alternatively, any of the above-described measurement devices may be configured such that the measurement unit can move in the thermostatic chamber 3 and can measure the state of cells in culture in each culture vessel noninvasively.

  The control device 2 is a normal computer including a processing unit including a central processing unit (CPU), a storage unit, and an input / output unit including a display 18 and a keyboard. The control device 2 operates the various programs 19 stored in the storage unit on the CPU as the processing unit using the various types of information stored in the database 20, thereby measuring the temperature adjustment unit 5 to the cell activity measurement. By controlling the apparatus 16, it is possible to control the culture environment in the thermostat 3 and to perform desired culture in the culture container 4 by examining the cell quality. The program 19 also includes a quality inspection program (quality inspection unit), and the user selects a menu screen on the display screen 17 that is one of the input / output units (user interface) of the control device 2 to perform the quality inspection. The unit can be activated. A series of cell tissue inspections are automatically performed by the quality inspection unit by inputting information necessary for the inspection. The user can select a menu on the display screen and appropriately display information on the inspection process and the inspection result. The display screen 17 may be provided with a GUI function so that the user can perform input / output operations by touching the screen.

FIG. 2 is a diagram showing an example of the program 19 of FIG. 1, and a quality inspection for executing a series of quality inspections in a plurality of stages using an epithelial cell sheet (when a barrier function is required) as a target tissue. It is an example of a program (quality inspection unit). The user who activates the quality inspection unit inputs data on the cell tissue to be measured, conditions / parameters related to the inspection, and the like from the display screen 17 (step S0). First, the quality inspection unit uses the cell observation device 12 or the impedance measurement device 13 to measure the impedance of the cell tissue to be measured with an alternating current and an alternating voltage at a high frequency (for example, 4000 Hz to several tens of thousands Hz), The state of cell proliferation is monitored (step S1). Next, this measured impedance is collated with a database of past data (step S2). FIG. 3 is a diagram showing an example of the database (table 201) used in step S2 of FIG. This table 201 gives the relationship between cell density and high frequency impedance. When the density is low (FIG. 3a), the impedance is lower than the threshold Z1, and when the density is in an appropriate range (FIG. 3b), the impedance is in the vicinity of the predetermined threshold Z1. Further, when the density is high (FIG. 3c), the impedance is high and shows a value sufficiently higher than the threshold value Z1. (It goes without saying that conductance may be used instead of impedance.)
FIG. 4 shows an example of a database (table 202) showing a state in which the high frequency impedance is increased and the time T during which the steady state is reached differs depending on the cell seeding density at the start of culture. When the seeding density is high (FIG. 4a), the impedance exceeds the threshold value Z2 at time t1 and further rises to reach a steady state. When the density is in an appropriate range (FIG. 4b), the impedance exceeds the value Z2 at time t2. On the other hand, when the sowing density is low (FIG. 4c), the time t3 until the impedance exceeds the threshold value Z2 is delayed.

  The quality inspection unit inspects whether there is a change in impedance Z in the cell tissue to be measured at a desired time (step S3). That is, the presence or absence of a change in impedance is determined from information in the database (tables 201 and 202). For example, the threshold value Z (= Z1 = Z2) is set, and if the impedance exceeds the threshold value Z by time t2, it is determined that there has been a change. Alternatively, the determination condition may be appropriately changed by the user according to the use of the cell tissue to be measured.

  Thus, the quality inspection unit can grasp the growth state of the cells by executing steps S1 to S3.

  If no impedance change is observed (NO in step S3), the cell activity of the cell tissue to be measured is then measured by the appropriate cell activity measuring device 16 (step S4). First, it collates with the database of past data (step S5), and the presence or absence of cell activity is inspected (step S6).

  A table 203 in FIG. 5 is a diagram showing an example of a database used in steps S5 and S6 in FIG. 2, and shows a relationship between cell activity and cell respiration activity that can be acquired when cells are measured with an electrochemical microscope. is there. When the cell activity is low, that is, when the cell is not in the growth phase state (FIG. 5a), the respiratory activity is lower than the threshold value (B1), and in the intermediate growth phase state (FIG. 5b), the respiratory activity is near the threshold value (B1). In addition, when the cell activity is high, that is, when the cell is actively proliferating (FIG. 5c), the respiratory activity is high and shows a value sufficiently higher than the threshold (B1). From the information in these databases (tables), it is determined whether or not cell activity is recognized (step S6 in FIG. 2). If the cell activity is lower than the threshold (B1) (NO in step S6), it is determined that the cell is dead or is in a state equivalent thereto, and the cell culture is stopped (step S7). When cell activity is recognized, culture | cultivation is continued (step S8) and step S1-3 is performed again. Noninvasive measurement techniques for cell activity include, but are not limited to, cell respiration activity measurement with an electrochemical microscope, cell viability measurement with laser polarization, and cell cycle measurement with a phase shift laser. In recent years, it has been clarified that the energy production mode differs between the undifferentiated state and the differentiated state of stem cells (undifferentiation: glycolytic pathway, differentiation: oxidative phosphorylation pathway). From this finding, an electrochemical microscope capable of examining the oxygen consumption of cells has the potential to examine the undifferentiated state of stem cells.

  When the impedance change is recognized in step S3 (Yes), the culture is continued. As the next step, the impedance of the cell tissue to be measured is measured with an alternating current and an alternating voltage at a low frequency (for example, several tens of Hz to 1,000 Hz), and the barrier function due to tight junction formation is monitored (step S9). Next, it collates with a database of past impedance data (step S10), and checks whether there is a change in impedance at a desired time (step S11).

  A table 204 in FIG. 6 is a diagram showing an example of a database used in steps S10 and S11 in FIG. 2, and shows a relationship between the barrier functionality and the low-frequency impedance. When there is no barrier function (FIG. 6a), the impedance is lower than the predetermined threshold value Z3, and when there is an appropriate barrier function (FIG. 6b), the impedance is near the threshold value Z3. When the barrier function is high (FIG. 6c), the impedance is high and shows a value larger than the threshold value Z3. Here again, as described above, the impedance rises and the time T until the steady state varies depending on the cell seeding density as shown in FIG. From the information in these databases (tables), it is determined whether or not a change in impedance is recognized at a desired time in step S11 of FIG.

  Thus, the function of the cell can be grasped by the quality inspection unit executing steps S9 to S11.

  If no change in impedance is observed in the cell tissue to be measured, the culture is stopped (step S12). On the other hand, when a change in impedance is recognized, the shape of the cell tissue to be measured is measured by the appropriate noninvasive cell / tissue shape measuring device 14 (step S13). It is checked against the database of past data (step S14), and the presence or absence of a desired cell / tissue shape is inspected (step S15).

  The table 205 in FIG. 7 is a diagram showing an example of the database used in steps S14 and S15 in FIG. 2, and shows the degree of stratification that can be obtained from cell / tissue shape analysis (for example, analysis by optical coherence tomography). It is. When the cell layer is 1 layer (Fig. 7a), the degree of layering is small and unsuitable as the tissue shape, and when the cell layer is several layers (4-5 layers or more) (Fig. 7c), it is optimal as the tissue shape. It is. When the cell layer is about two layers (FIG. 7b), it is necessary to determine whether or not the shape is a desired shape depending on the application. Cell layer shape determination conditions (threshold values) are set in advance.

  Step S15 in FIG. 2 is determined from information in these databases (tables) and a preset threshold value. If the cell tissue to be measured is not in the desired shape, the culture is continued assuming that the threshold has not been reached (step S16), and steps S13 to S16 are performed again. If the desired shape is equal to or greater than the threshold, it is determined that the cell tissue is transplantable, and the culture is terminated. Non-invasive measurement of cell / tissue shape includes measurement of tissue cross-section thickness using optical coherence tomography, non-contact AFM (atomic force microscope) and non-linear optical methods (Raman microscope and CARS (coherent anti-virus)). The cell surface fine structure measurement by stroke Raman spectroscopy, SHG (second harmonic generation), SFG (sum frequency generation), etc.) is conceivable, but is not limited thereto.

  In addition, regarding the frequency at the time of the above-mentioned impedance measurement, it described based on the following general knowledge, and is not limited to this. In general, at low frequencies, the current bypasses cells and flows through the gaps between epithelial cells, so the impedance value obtained by measurement reflects the state of tight junctions between cells, and at high frequencies, It is known that the current does not bypass the cell, and the impedance value obtained by measurement reflects the state of cell proliferation (see Non-Patent Document 2).

  As described above, according to the present embodiment, a series of quality inspection steps necessary for multipoint observation of cells / tissues can be performed non-invasively and automatically for the cell tissue to be measured. That is, if the cell growth status (concentration, cell activity) is grasped and it satisfies the predetermined condition, then the cell function (barrier function) is grasped and it satisfies the predetermined condition. Then, the presence / absence (polymerization degree) of a desired cell / tissue shape can be observed non-invasively and automatically.

FIG. 8 shows an example of the display screen 17 on the display device 18 when performing the culture process of the cell culture device according to the first embodiment. In FIG. 8, on the display screen 17, the cell observation result 21 and the impedance measurement result 22 are displayed in the image display area. In the display screen 17, an environmental temperature, humidity, CO ₂ concentration, and temperature / humidity / concentration indicating a concentration of O ₂ in the thermostatic chamber 3 and a culture time display area 23 are displayed. Furthermore, a button 24 for selecting an automatic sequence or a manual sequence of the culture process, a button 25 for selecting a culture medium exchange, and a button 26 for selecting cell observation are displayed. Further, a button 27 for selecting impedance measurement (high frequency), a button 28 for selecting impedance measurement (low frequency), a button 29 for selecting cell activity measurement, a button 30 for selecting cell / tissue shape measurement, and an action potential measurement mode. A selection button 31 is displayed. For example, when 24 tissues are cultured as in the display area 40 of the configuration of the culture container, it is possible to display data on one tissue or data on a plurality of tissues. In this example, the user operates the menu screen of the display screen 17 to display the cell observation image 21 and the impedance (high frequency) measurement result 22 on the screen for the culture vessel A-1. The user sets the conditions necessary for multipoint observation of cells / tissues by the quality inspection unit for the cell tissue to be measured, and the data that he / she wants to view regarding the status and results of a series of quality inspections that are automatically processed Can be displayed and observed in real time. It should be noted that it is also possible for the user to change parameters and determination conditions in accordance with the state of the cell tissue to be measured displayed on the display screen 17 during the examination.

  According to the present embodiment, a series of quality inspections for multipoint observation of cells / tissues to be transplanted can be performed non-invasively and automatically, and safe and secure regenerative medicine can be realized.

  Next, a second embodiment of the present invention will be described with reference to FIGS. FIG. 9 shows an example of a quality inspection program (quality inspection unit) in which the target tissue is a cell sheet that generates spontaneous action potentials such as myocardium, myoblast, and nerve.

  The user who has started the quality inspection unit inputs data on cell tissue to be measured, conditions / parameters related to inspection, and the like from the display screen 17. Subsequently, steps S1 to S8 are the same as those in the first embodiment. If an impedance change is recognized in step S3, the culture is continued. As the next step, the action potential is measured by the cell observation device 12 and the action potential measuring device 15 (step S17), checked against a database of past data (step S18), and inspected whether a desired action potential is obtained. (Step S19).

  FIG. 10 is a diagram showing an example of the database 206 used in steps S18 and S19 of FIG. 9, and shows the relationship between action potentials and cell membrane potential values. When the cell-specific functions and activities are low (10a), the membrane potential changes when depolarizing after stimulation is small and smaller than the threshold value V1. When cell-specific functions and activities become moderate (FIG. 10b), the membrane potential change exceeds the predetermined threshold value V1, and when cell-specific functions and activities are high (FIG. 10c), depolarization occurs after stimulation. The change in the membrane potential at the time is large and shows a value sufficiently larger than the threshold value V1. From these databases, it is determined whether or not there is an action potential in step S19 in FIG.

  If no action potential is observed, the culture is stopped (step S20). If an action potential is recognized, the tissue shape of the cell is measured by an appropriate noninvasive tissue shape measuring means (step S13). Further, in the same manner as in the first embodiment, it is checked against the database of past data (step S14), and the presence or absence of a desired cell / tissue shape is inspected (step S15). When it is not a desired shape, culture | cultivation is continued (step S16) and step S13-16 is performed again. When the desired shape is obtained, the culture is terminated. Non-invasive measures of action potential include measurement of potential changes in the vicinity of cells using a multi-electrode array, measurement of magnetic field associated with action potential generation using a SQUID (superconducting quantum interference device) magnetometer, and action potential using an electrochemical microscope. Although potential change measurement in the vicinity of cells can be considered, it is not limited to this. In addition, the culture dish with electrodes is often opaque and becomes an obstacle when observing cells. In this case, the electrode shape may be devised or a semitransparent semiconductor may be used.

  Non-invasive measurement of cell shape includes tissue cross-sectional thickness measurement by optical coherence tomography, non-contact high-speed AFM, nonlinear optical technique (Raman microscope, CARS (coherent anti-stroke Raman spectroscopy), SHG ( Cell surface fine structure measurement by second harmonic generation), SFG (sum frequency generation, etc.) is conceivable, but is not limited to this. The display screen 17 shown in FIG. 8 is also applicable to the second embodiment.

  Also in this embodiment, a series of quality inspection steps necessary for quality evaluation using a plurality of cell / tissue indices can be performed non-invasively and automatically for the cell tissue to be measured. That is, if the growth status (concentration, cell activity) of a cell is grasped and it satisfies a predetermined condition, then the cell-specific function and activity are grasped and the condition is satisfied. In this case, next, the presence / absence (polymerization degree) of a desired cell / tissue shape can be observed non-invasively and automatically. That is, a series of quality inspections for quality evaluation using a plurality of indexes of cells / tissues to be transplanted can be performed non-invasively and automatically, and safe and secure regenerative medicine can be realized.

  Next, a third embodiment of the present invention will be described with reference to the drawings. FIG. 11 shows an example of a quality inspection program (quality inspection unit) in which a cell sheet other than the tissue targeted in Examples 1 and 2 is the target tissue. The user who has started the quality inspection unit inputs data on cell tissue to be measured, conditions / parameters related to inspection, and the like from the display screen 17. Subsequently, steps S1 to S8 are the same as in the first and second embodiments. If an impedance change is recognized in step S3, the culture is continued. As the next step, the tissue shape is measured by a suitable non-invasive cell / tissue shape measuring means (step S13). It is checked against the database of past data (step S14), and the presence or absence of a desired cell / tissue shape is inspected (step S15). When it is not a desired shape, culture | cultivation is continued (step S16) and step S13-16 is performed again. When the desired shape is obtained, the culture is terminated. The display screen 17 shown in FIG. 8 is also applicable to the third embodiment.

  Also in this embodiment, a series of quality inspections for quality evaluation using a plurality of indices of cells / tissues to be transplanted can be performed non-invasively and automatically, and safe and secure regenerative medicine can be realized.

  FIG. 12 is a diagram showing another example of the display screen used in the first to third embodiments according to the fourth embodiment of the present invention. Unlike the example of FIG. 8, the data to be displayed can display not only a single measurement result but also the results obtained by a plurality of types of measurement. The data that the user wants to browse differs depending on the culture target and the culture elapsed time, and a display method using all possible combinations may be used. In the example of FIG. 12, two types of data are displayed: an impedance (high frequency) measurement result 22 and an impedance (low frequency) measurement result 33.

  The display screen does not need to be directly connected to the control device, and may be displayed on a terminal connected by wireless communication such as a tablet terminal or a smartphone. By using such a wireless communication terminal, the cell culture apparatus can be remotely operated and the operation content can be confirmed, which is beneficial to the apparatus operator. In addition, it is possible to reduce human errors by instructing a device operator having a low skill level to instruct the device operation method and displaying the screen so as to prompt an operation to be performed.

  In addition, this invention is not limited to an above-described Example, It is applicable to all cell types. Further, the above-described embodiments have been described in detail for easy understanding of the present invention, and are not necessarily limited to those having all the configurations described. Further, a part of the configuration of one embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of one embodiment. Further, it is possible to add, delete, and replace other configurations for a part of the configuration of each embodiment.

  Furthermore, each configuration, function, processing unit, processing means, and the like including the quality inspection unit may be realized by dedicated hardware by designing a part or all of them with, for example, an integrated circuit. Information such as a control program that realizes each function of the quality inspection unit is stored not only in a memory as a storage unit but also in a storage device such as a hard disk or an SSD (Solid State Drive), or a recording medium such as an IC (Integrated Circuit) card. I can leave.

DESCRIPTION OF SYMBOLS 1 ... Cell culture apparatus 2 ... Control apparatus 3 ... Constant temperature bath 4 ... Culture container 13 ... Impedance measurement apparatus 14 ... Cell and tissue shape measurement apparatus 15 ... Action potential measurement apparatus 16 ... Cell activity measurement apparatus 17 ... Display screen 18 ... Display 19 ... Quality inspection program (Quality inspection unit)
20 ... Database 21 ... Cell observation result 22 ... Impedance measurement result 201 ... Table 202 ... Table 203 ... Table 204 ... Table 205 ... Table 206 ... Table

Claims (15)

A cellular tissue examination system for non-invasive examination of cellular tissue in multiple stages ,
A measuring device used for non-invasive examination in the plurality of stages ;
A quality inspection unit for controlling non-invasive inspection of the cell tissue to be measured using the measuring device in the plurality of stages ;
A database for holding information on non-invasive examinations in the plurality of stages ,
The non-invasive examination in the plurality of stages includes at least a grasping examination of the growth state of the cells,
The impedance of the measurement target tissue is measured, the measurement result is compared with a predetermined first threshold value, and the next inspection is performed when the threshold value is equal to or greater than the first threshold value. A cell tissue examination system characterized by changing the contents of. In claim 1,
The non-invasive examination in the plurality of stages includes an examination on the function of the cell and an examination on the tissue shape of the cell,
When the measurement result is equal to or greater than the first threshold, the cell tissue inspection system is characterized by measuring the function of the cell or the tissue shape of the cell. In claim 2,
In order to grasp and check the growth status of the cells, the high frequency impedance of the measurement target tissue is measured, and the measurement result is compared with the first threshold value,
If the measurement result is greater than or equal to the first threshold value, the low-frequency impedance of the cell tissue to be measured is measured for a test relating to the function of the cell, and the measurement result is compared with a predetermined third threshold value Make a decision,
When the measurement result of the low-frequency impedance is equal to or greater than the third threshold, the tissue shape of the measurement target cell is measured, the measurement result is compared with a predetermined second threshold, and the second A cell tissue examination system, wherein the next processing content is changed depending on whether the threshold is equal to or greater than the threshold. In claim 2,
If it is equal to or greater than the first threshold, the action potential of the cell is measured, and the measurement result is compared with a predetermined fifth threshold,
When the action potential measurement result is equal to or greater than the fifth threshold value, the tissue shape of the cell is measured, and the measurement result is compared with a predetermined second threshold value. And a cell tissue examination system characterized in that the next processing content is changed depending on whether or not the threshold is satisfied. In claim 1,
The non-invasive examination in the plurality of stages includes an examination regarding the tissue shape of the cell,
If it is greater than or equal to the first threshold, the tissue shape of the cell is measured, the measurement result is compared with a predetermined second threshold, and if the threshold is greater than or equal to the second threshold, the threshold is met. A cell tissue examination system characterized in that the next processing content is changed when there is no. In any of claims 3 to 5,
When the first threshold value is not reached, the cell activity of the cell tissue to be measured is measured, the measurement result of the cell activity is compared with the fourth threshold value, and the measurement result of the cell activity is If the fourth threshold value or more, continue the culture of the measurement target cell tissue, if the measurement result of the cell activity is less than the fourth threshold, stop the cell culture,
When the second threshold or more, it is determined that the measurement target tissue is a transplantable cell, and when the measurement result is less than the second threshold, the tissue shape is measured again, A cellular tissue examination system, which performs a comparison determination with the second threshold value. In claim 1,
A cell tissue examination system comprising a display that functions as a user interface and displays a measurement target of a non-invasive examination in the plurality of stages and at least one positioning result. A culture vessel for cell culture;
A thermostat for cell culture in which the culture vessel is installed;
A control device for controlling the culture environment of the culture vessel;
A cell observation device;
An impedance measuring device;
A cell / tissue morphology measuring device;
A cell activity measuring device;
A cell culture device comprising:
The controller is
A quality inspection unit for controlling a multi-stage non-invasive cell inspection using each measuring device,
The multi-stage non-invasive cell examination includes grasping the growth state of cells and measuring the tissue shape,
The quality inspection unit is
For grasping and checking the growth state of the cell, the impedance of the cellular tissue is measured by the impedance measuring device, the measurement result is compared with a predetermined first threshold value, and is equal to or higher than the first threshold value. A cell culture device characterized in that the contents of the next examination are changed depending on the case and the case where the threshold is not met. In claim 8,
For the purpose of grasping and examining the growth state of the cells, the high-frequency impedance of the cellular tissue is measured by the impedance measuring device,
The cell culture device characterized in that when the impedance is equal to or higher than the first threshold, the low-frequency impedance of the cell is measured by the impedance measurement device. In claim 9,
Equipped with action potential measuring device,
For the purpose of grasping and examining the growth state of the cells, the high-frequency impedance of the cellular tissue is measured by the impedance measuring device,
The cell culture device, wherein the action potential of the cell is measured when the first threshold value or more. In claim 8,
For the purpose of grasping and examining the growth state of the cells, the high-frequency impedance of the cellular tissue is measured by the impedance measuring device,
The cell culture device, wherein the tissue shape is measured by the cell / tissue morphology measuring device when the first threshold value or more. In claim 8,
A cell culture device that functions as a user interface and includes a display that displays at least one measurement object and the positioning result of each of the plurality of non-invasive cell tests. A method for non-invasively examining a tissue of a cell in multiple stages using a cellular tissue examination system,
The cell tissue examination system comprises:
A measuring device used for non-invasive examination in the plurality of stages ;
A quality inspection unit for controlling the inspection of the cell tissue to be measured using the measuring device;
A database holding information on the inspection,
The non-invasive examination in the plurality of stages includes grasping the growth state of cells and measuring the tissue shape,
For grasping and examining the growth state of the cells, the impedance of the measurement target tissue is measured,
A cellular tissue characterized in that the measurement result is compared with a predetermined first threshold value, and the content of the next examination is changed between the case where the measurement result is equal to or more than the first threshold value and the case where the measurement result is less than the threshold value. Inspection method. In claim 13,
If the measurement result is greater than or equal to the first threshold, measure the tissue shape of the cell or the function of the cell,
When the first threshold value is not satisfied, the measurement of the impedance of the measurement target cell tissue is continued and the comparison determination is performed again after continuing the culture of the measurement target cell tissue. . In claim 13,
If the measurement result is greater than or equal to the first threshold, measure the action potential of the cell,
When the first threshold value is not satisfied, the measurement of the impedance of the measurement target cell tissue is continued and the comparison determination is performed again after continuing the culture of the measurement target cell tissue. .

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